Project Summary Applikate Technologies has developed Clearing Histology with Multiphoton Microscopy (CHiMP), a novel tissue processing and imaging protocol that provides direct-to-digital images from intact pathology tissue specimens, such as biopsies, without the need for wax embedding, slicing, or scanning of slides. The application of this technology to renal diagnosis is expected to improve considerably our ability to render accurate, cost-effective, and timely diagnoses in patients who suffer from a broad range of kidney problems. It will also expand our ability to study and understand kidney disorders by providing otherwise inaccessible data which can be analyzed with emerging computer analysis tools. The aim of this direct-to-Phase II SBIR proposal is to modify the hardware and software components of CHiMP for application to kidney biopsies and to validate CHiMP's non-inferiority for renal diagnosis compared to standard protocols and its integration into the renal pathology workflow in a clinical setting. CHiMP has many advantages for general histology, including: H&E-like images familiar to pathologists but with superior image quality, sub-micron effective thickness, and without artifacts common to slide preparation; non- destructive imaging that preserves intact tissue for ancillary molecular analysis and immunohistochemistry; rapid turnaround, with images ready within a few hours of receiving tissue; greatly reduced labor costs; true 3D prospective; and digitized images for remote consultation and machine vision. NIDDK has identified access to human kidney biopsies as an important bottle neck for progress in development of precision medicine for acute and chronic kidney disease. CHiMP directly addresses this by preserving tissue and increasing DNA yields. In addition, this proposal will extend CHiMP's imaging to include second harmonic generation from collagen, thereby enabling evaluation and quantitation of fibrosis, a key factor in renal health and applicable to other conditions such as liver disease. The specific aims of this proposal are to: 1) modify the CHiMP microscope and tissue cassettes to enable second harmonic generation imaging of collagen in kidney biopsies, 2) collect renal biopsy specimens in a clinical setting and evaluate integration into the clinical workflow and 3) add renal specific visualization features to Stackstreamer, our 3D visualization software, and demonstrate the utility of CHiMP for diagnosis. Successful completion of these aims will set the stage for a Phase III transition to regulatory approval and provide a proven platform for early implementation in select hospitals.